Nature Biotechnology
21, 652 - 659 (2003)
Published online: 5 May 2003; Corrected online: 30 May 2003 | doi:10.1038/nbt822
There is a Corrigenda (July 2003) associated with this Article.
High-throughput engineering of the mouse genome coupled with
high-resolution expression analysisDavid M Valenzuela1, Andrew J Murphy1, 3, David Frendewey1, 3, Nicholas W Gale1, 3, Aris N Economides1, 3, Wojtek Auerbach1, William T Poueymirou1, Niels C Adams1, Jose Rojas1, Jason Yasenchak1, Rostislav Chernomorsky1, Marylene Boucher1, Andrea L Elsasser1, Lakeisha Esau1, Jenny Zheng1, Jennifer A Griffiths1, Xiaorong Wang1, Hong Su1, Yingzi Xue1, Melissa G Dominguez1, Irene Noguera1, Richard Torres1, Lynn E Macdonald1, A Francis Stewart2, Thomas M DeChiara1
& George D Yancopoulos11
Regeneron Pharmaceuticals, Inc., 777
Old Saw Mill Road, Tarrytown, New York
10591, USA. 2
BIOTEC, Technische Universitat Dresden,
Dresden, Germany 01307. 3
These authors contributed equally to this work.
Correspondence should be addressed to George D Yancopoulos velocigene@regeneron.comOne of the most effective approaches for determining gene function
involves engineering mice with mutations or deletions in endogenous genes of
interest. Historically, this approach has been limited by the difficulty and
time required to generate such mice. We describe the development of a
high-throughput and largely automated process, termed VelociGene, that uses
targeting vectors based on bacterial artificial chromosomes (BACs). VelociGene
permits genetic alteration with nucleotide precision, is not limited by the
size of desired deletions, does not depend on isogenicity or on
positive−negative selection, and can precisely replace the gene of
interest with a reporter that allows for high-resolution localization of
target-gene expression. We describe custom genetic alterations for hundreds of
genes, corresponding to about 0.5−1.0% of the entire genome. We also
provide dozens of informative expression patterns involving cells in the
nervous system, immune system, vasculature, skeleton, fat and other
tissues.*Note: In the author list of the AOP version of this article, the name
of author Rostislav Chernomorsky was misspelled Rostislav Chernomorski. This
has been corrected in the online and print versions of the article.
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